青海省海晏县线叶嵩草内生细菌的生物功能鉴定及测定
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摘要
对线叶嵩草(Kobresia capillifolia)内生细菌部分生物功能进行鉴定及测定;采用组织分离法从青藏高原青海省海西线叶嵩草根部分离出10株内生细菌。结果表明,10株内生细菌均能固氮,2G1和2G10能够产IAA;其中2G2、2G3、2G4、2G5、2G6、2G7、2G8和2G9对指示菌均有拮抗作用,对马铃薯炭疽菌Colletotrlchum coccodes的拮抗作用最强,抑菌率达88.58%。通过培养性状和形态特征,结合16SrDNA序列相似性分析,将2G1和2G10分别鉴定为苏云金芽孢杆菌Bacillus thuringiensis和Bacillus simpix,2G2和2G9鉴定为亲甲基醇芽孢杆菌Bacillus methylotrophicus,2G3和2G6鉴定为解淀粉芽孢杆菌Bacillus amyloliquefaciens,2G4和2G5鉴定为云南苏铁芽孢杆菌Bacillus siamensis,2G7和2G8鉴定为枯草芽孢杆菌Bacillus subtilis。该研究结果为青藏高原极端生境牧草根部内生细菌的多样性及功能菌株的开发提供依据。
To identify and determine the bioactivities of endophytic bacteria in Kobrecia capillifolia,ten isolates of endophytic bacteria were isolated from the roots of Kobresia capillifolia from Haixi Qinghai Province,Qinghai-Tibet Plateau by using tissue culture method.The results showed that all 10 endophytic bacteria were able to fix nitrogen,and 2G1 and 2G10 could produce IAA.Among them,2G2,2G3,2G4,2G5,2G6,2G7,2G8 and 2G9 all antagonized the indicator bacteria and antagonized the potato anthracnose strongest,antibacterial rate of 88.58%.By tracing the traits and morphological characteristics,combining with 16 SrDNA sequence similarity analysis,2G1 and 2G10 were identified as Bacillus thuringiensis and Bacillus simpixrespectively,while 2G2 and 2G9 were identified as Bacillus methylotrophicus,2G3 and 2G6 were identified as Bacillus amyloliquefaciens,2G4 and 2G5 were identified as Bacillus siamensis,2G7 and 2G8 were identified as Bacillus subtilis.The results provide a basis for the diversity of endophytic bacteria and the development of functional strains in the extreme habitats of the Qinghai-Tibet Plateau.
To identify and determine the bioactivities of endophytic bacteria in Kobrecia capillifolia,ten isolates of endophytic bacteria were isolated from the roots of Kobresia capillifolia from Haixi Qinghai Province,Qinghai-Tibet Plateau by using tissue culture method.The results showed that all 10 endophytic bacteria were able to fix nitrogen,and 2G1 and 2G10 could produce IAA.Among them,2G2,2G3,2G4,2G5,2G6,2G7,2G8 and 2G9 all antagonized the indicator bacteria and antagonized the potato anthracnose strongest,antibacterial rate of 88.58%.By tracing the traits and morphological characteristics,combining with 16 SrDNA sequence similarity analysis,2G1 and 2G10 were identified as Bacillus thuringiensis and Bacillus simpixrespectively,while 2G2 and 2G9 were identified as Bacillus methylotrophicus,2G3 and 2G6 were identified as Bacillus amyloliquefaciens,2G4 and 2G5 were identified as Bacillus siamensis,2G7 and 2G8 were identified as Bacillus subtilis.The results provide a basis for the diversity of endophytic bacteria and the development of functional strains in the extreme habitats of the Qinghai-Tibet Plateau.
引文
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[27]张芳芳,宋希强,朱国鹏.不同生境下五唇兰根部可培养内生细菌多样性研究[J].植物科学学报,2016,34(1):135-142
[2]文才艺,吴元华,田秀玲.植物内生菌研究进展及其存在的问题[J].生态学杂志,2004,23(2):86-91
[3]喻江,于镇华,刘晓冰,等.植物根组织内生细菌多样性及其促生作用[J].中国农学通报,2015,31(13):169-175
[4]林吉恒,王睿之,马风兰,等.植物益生菌对植物的益生效应及其应用[J].中国生物防治,2010,26:100-105
[5]戴传超,余伯阳,赵玉婷,等.大戟内生菌的抑菌活性及其与宿主相关性研究[J].应用生态学报,2015,16(7):1290-1294
[6]王娜,陶伟,陈双林,等.植物内生细菌在棉花体内的定殖动态及对棉花黄萎病的生物防治效果[J].植物保护学报,2016,43(2):207-214
[7]杜晓宁,徐惠娟,黄盼盼,等.宁夏枸杞内生细菌的多样性及其抑菌活性研究[J].微生物学通报,2015,42(9):1779-1787
[8]蔡学清,陈炜,林娜,等.内生细菌在荔枝体内的定殖及其防病保鲜功能[J].应用生态学,2011,22(8):2140-2146
[9]李峰,徐大勇,王光利,等.一株拮抗赤霉病的小麦内生细菌的筛选和抑菌活性[J].生态学杂志,2011,30(8):1738-1743
[10]崔月贞,万志文,冯疆蓉,等.东祁连山高寒草地优势牧草内生细菌产IAA能力的研究[J].草地学报,2016,24(3):618-623
[11]高晓星,满百膺,陈秀蓉,等.东祁连山线叶嵩草内生细菌X4的产吲哚乙酸、解磷、抗菌和耐盐特性研究及分子鉴定[J].草业学报,2013,22(4):137-146
[12]方中达.植病研究法[M].北京:中国农业出版社,1996:122-154
[13]徐爱芳,武永秀,刘莹.春兰根内生解有机磷细菌的筛选[J].安徽农业科学,2013,41(4):1515-1516,1529
[14]Xu L Q,Yao T,Yang J J.Porperty of associative nitrogen-fixing bacteria producing IAA and its promoting growth of oat[J].Grassland and Turf,2005,(4):25-29
[15]满百膺,陈秀蓉,李振东.高寒牧草内生细菌分离培养条件的优化[J].草原与草坪,2008,(5):27-30
[16]Vetrivelkalai P,Sivakumar M,Jonathan E I.Biocontrol potential of endophytice bacteria on Meloidogyne incognita and itseffect on plant growth in bhendi[J].Journal of Biopesticides,2010,3(2):452-457
[17]Fiore S D,Gallo M.Endophytic bacteria:Their possible role in the Host Plant A-Zospirollum VI and related microorganisms[J].Germany:Springer-Verlag,1995,169-187
[18]胡青平,徐建国,刘会龙,等.番茄茎内生细菌的分离鉴定及青枯病拮抗菌的筛选[J].西北植物学报,2006,26(10):2039-2043
[19]崔月贞,杨小利,杨成德,等.拮抗马铃薯晚疫病菌的高寒草地牧草内生细菌的鉴定及其生物功能测定[J].植物保护学报,2016,43(5):789-795
[20]Glickmann E,Dessaux Y.A critical examination of the specificity of compounds produced by phytopathogenic bacteria[J].Applied and Environmental Microbiology,1995,619(2):793-796
[21]东秀珠,蔡妙英.常见细菌系统鉴定手册[M].北京:中国科学出版社,2001
[22]Mou X M,Li X G,Zhao N N,et al.Tibetan sedges sequester more carbon belowground than grasses a 13Clabeling study[J].Plant and Soil,2018,426(1-2):287-298
[23]杨成德,王玉琴,陈秀蓉,等.2种嵩草属牧草休眠期内生细菌多样性研究[J].草业学报,2016,25(8):136-144
[24]Forchetti G,Masciarelli O,Izaguirre M,et al.Endophytic bacteria improve seedling growth of sunflower under water stress,produce salicylic acid,and inhibit growth of pathogenic fungi[J].Current Microbiology,2010,61:485-493
[25]Zhang G X,Peng G X,Wang E T.Diverse endophytic nitrogen-fixing bacteria isolated from wild rice Oryza rufipogon and description of Phytobacter diazotrophicus gen.nov.sp.nov.[J].Archives of Microbiology,2008,189:431-439
[26]姜红霞,杨成德,陈秀蓉,等.甘肃省马铃薯坏疽病鉴定及其病原生物学特性研究[J].草业学报,2013,22(2):123-131
[27]张芳芳,宋希强,朱国鹏.不同生境下五唇兰根部可培养内生细菌多样性研究[J].植物科学学报,2016,34(1):135-142